Composition for enhancing immune function and application

Abstract

The invention discloses a composition for enhancing an immune function and application, and belongs to the technical field of pharmaceutical preparations. According to the invention, 1.4 to 1.8 parts of total saponins of ginseng, 12 to 16 parts of L-arginine hydrochloride, 18 to 22 parts of L-glutamine, 0.18 to 0.22 part of magnesium vitamin C phosphate, 0.02 to 0.06 part of zinc gluconate, 2 to 6 parts of DHA, 0.6 to 1.0 part of EPA and 0.000008 to 0.000012 part of cholecalciferol are selected to prepare the composition for enhancing the immune function, and according to an animal efficacy evaluation result, the composition can be used for preparing the composition for enhancing the immune function. The thymus index, the macrophage phagocytic ability and the NK cell activity of mice with low immunologic functions caused by cyclophosphamide can be improved, so that the body immunologic function is enhanced.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and in particular to a composition for enhancing immune function and its application. Background Art

[0002] Immune function is an important physiological function of the body to resist disease, block the invasion of pathogens, and maintain the stability of the body's internal environment. Factors such as unbalanced diet, mental stress, irregular work and rest, lack of exercise, excessive fatigue, closed environment, radiation exposure, etc. can all lead to damaged or low immune system function, which in turn makes the body extremely susceptible to infection by pathogens and has an adverse effect on the body's health.

[0003] In the prior art, drugs used to enhance the body's immunity include pidotimod, but it often causes damage to the digestive system, skin and appendages, and nervous system. Transfer factor usually also causes skin reactions, gastrointestinal reactions, and partial nervous system reactions. Thymosin for injection can cause allergic reactions. Excessive intake of vitamins can lead to poisoning or other health problems. BCG can cause local redness, swelling, suppuration, etc. Human immunoglobulin does not last long, is generally a blood product, and has certain risks and may cause dependence. Levamisole may cause allergic reactions, nausea and vomiting, etc.

[0004] Based on the above problems existing in the prior art, the present invention urgently needs to prepare a pharmaceutical composition for enhancing the immune function of the body that overcomes the above technical defects. Summary of the invention

[0005] Based on the above technical problems to be solved by the present invention, the present invention proposes a composition for enhancing immune function and its application, wherein the composition comprises total ginsenosides of Panax ginseng, L-arginine hydrochloride, L-glutamine, magnesium vitamin C phosphate, zinc gluconate, DHA, EPA, and cholecalciferol.

[0006] One of the purposes of the present invention is to provide a composition for enhancing immune function, wherein the components of the composition for enhancing immune function include, by weight:

[0007] 1.4 to 1.8 parts of total ginsenosides, 12 to 16 parts of L-arginine hydrochloride, 18 to 22 parts of L-glutamine, 0.18 to 0.22 parts of magnesium ascorbyl phosphate, 0.02 to 0.06 parts of zinc gluconate, 2 to 6 parts of DHA, 0.6 to 1.0 parts of EPA, and 0.000008 to 0.000012 parts of cholecalciferol.

[0008] Furthermore, the total ginsenosides are 1.6 parts, the L-arginine hydrochloride is 14 parts, the L-glutamine is 20 parts, the magnesium ascorbyl phosphate is 0.2 parts, the zinc gluconate is 0.04 parts, the DHA is 4 parts, the EPA is 0.8 parts, and the cholecalciferol is 0.00001 parts.

[0009] The second object of the present invention is to provide a preparation method of a composition for enhancing immune function, the preparation method comprising:

[0010] The ginsenosides, L-arginine hydrochloride, L-glutamine, magnesium ascorbyl phosphate, zinc gluconate, DHA, EPA and cholecalciferol are uniformly mixed.

[0011] The third object of the present invention is to provide a product consisting of a composition for enhancing immune function and one or more additives.

[0012] Furthermore, the product is a pharmaceutical composition, a health care product composition or a food composition.

[0013] Furthermore, the product is formulated into tablets, capsules or granules.

[0014] The third object of the present invention is to provide a method for preparing a pharmaceutical composition for enhancing the immune function of a subject.

[0015] Furthermore, the composition for enhancing immune function can regulate cellular immune function;

[0016] The composition for enhancing immune function can enhance humoral immune function;

[0017] The composition for enhancing immune function can enhance the phagocytic function of monocytes and macrophages;

[0018] The composition for enhancing immune function can enhance NK cell activity.

[0019] The fourth object of the present invention is to provide an application in the preparation of a product for enhancing immune function.

[0020] Furthermore, the product for enhancing immune function includes protein powder, multivitamins and aromatherapy.

[0021] Compared with the prior art, the present invention provides a composition and application for enhancing immune function, which has the following beneficial effects:

[0022] 1. According to the results of animal efficacy evaluation, the composition proposed by the present invention can improve the thymus index, macrophage phagocytic ability and NK cell activity of immunosuppressed mice induced by cyclophosphamide, thereby enhancing the body's immune function.

[0023] 2. The composition proposed by the present invention, based on the role of each main component in enhancing the activity and function of immune cells, regulates immune response and strengthens the immune barrier. The synergistic effect between the components promotes the body to play a comprehensive and effective role in immune regulation. The synergistic effect in immune regulation includes that total ginsenosides, L-arginine hydrochloride and L-glutamine promote the proliferation and activation of immune cells through different mechanisms; magnesium vitamin C phosphate and zinc gluconate enhance the function of immune cells by anti-oxidation and supplementation of trace elements; DHA, EPA and cholecalciferol enhance the body's immunity by regulating immune cell signal transduction and immune response.

[0024] 3. The composition prepared by the present invention has a better synergistic effect on improving human immune function compared with the use of total ginsenosides alone. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The effect of mouse spleen T lymphocyte transformation on the embodiment of the present invention is shown (x±

[0026] s, n = 10, mg / g); * compared with group C, P < 0.05, # compared with group M, P < 0.05;

[0027] Figure 2 The present invention shows a method for expressing CD4+ in mouse peripheral blood lymphocytes. 3+ CD 4+ and CD 3+ CD 8+ T cell expression (x±s, n=3); *Compared with group C, P<0.05, #Compared with group M, P<0.05;

[0028] Figure 3 The immunoglobulin content of mouse serum in an embodiment of the present invention is shown (x±s, n=10, mg / g); * compared with group C, P<0.05, # compared with group M, P<0.05. DETAILED DESCRIPTION

[0029] The exemplary embodiments of the present application will be described in more detail below with reference to the accompanying drawings. Although the exemplary embodiments of the present application are shown in the accompanying drawings, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided in order to enable a more thorough understanding of the present application and to fully convey the scope of the present application to those skilled in the art.

[0030] In the following examples, unless otherwise specified, the technical means adopted are conventional means well known to those skilled in the art, and the reagents and materials in the present invention are obtained from the market or other public channels.

[0031] The experimental materials and equipment involved in the present invention mainly include but are not limited to:

[0032] Reagents:

[0033] Ginseng total saponins (≥80%, root extract), Shanghai Yuanye Biotechnology Co., Ltd.; L-arginine hydrochloride (≥99%), Aladdin Biochemical Technology Co., Ltd.; L-glutamine (≥99%), Aladdin Biochemical Technology Co., Ltd.; Vitamin C phosphate magnesium (≥99%), Hongxin Biotechnology Co., Ltd.; zinc gluconate (≥99%), Jiangsu Jiayuancheng Biotechnology Co., Ltd.; DHA algae oil (39.6%), Newmans Holdings Co., Ltd.; EPA fish oil (98.5%), NuHaimINC; cholecalciferol (≥99%), Aladdin Biochemical Technology Co., Ltd.;

[0034] instrument:

[0035] Low-temperature high-speed centrifuge, 5810R, Eppendorf Innovation Company; electronic balance, ME204 / 02, METTLER TOLEDO; upright microscope, DM4 B, Leica Microsystems; super microplate reader, uQuantTM200-999, Bio Tec.

[0036] The composition and application of the present invention for enhancing immune function comprises ginsenosides, L-arginine hydrochloride, L-glutamine, magnesium vitamin C phosphate, zinc gluconate, DHA, EPA, and cholecalciferol. Based on the composition of the present invention, its immune synergistic effects mainly include:

[0037] Based on the role of each main component in enhancing the activity and function of immune cells, regulating immune response and strengthening immune barrier, the synergistic effect between the components promotes the body to play a comprehensive and effective role in immune regulation. The synergistic effect in immune regulation includes ginsenosides, L-arginine salts and L-glutamine promoting the proliferation and activation of immune cells through different mechanisms; magnesium vitamin C phosphate and zinc gluconate enhance the function of immune cells through anti-oxidation and supplementation of trace elements; DHA, EPA and cholecalciferol enhance the body's immunity by regulating immune cell signal transduction and immune response. Among them,

[0038] Ginseng total saponins: can enhance the number and activity of immune cells such as T cells and NK cells; ginsenoside Rh2 can promote the expression of co-stimulatory molecule MHCII on the surface of immune molecule dendritic cells, promote the secretion of cytokine interleukin 2 and TNF-α tumor necrosis factor in serum, and thus enhance the function of immune organs such as spleen and thymus.

[0039] L-arginine hydrochloride: It is the main substrate for the synthesis of nitric oxide. NO is an important biologically active molecule. L-arginine salt promotes the synthesis of NO, inhibits the production and release of inflammatory cytokines, and reduces the inflammatory response of tissues; enhances the phagocytosis, killing and proliferation capabilities of immune cells such as macrophages, NK cells and T cells; balances the activity of Th1 and Th2 cells; supports protein synthesis and immune cell repair, and maintains acid-base balance and a stable immune environment.

[0040] L-glutamine: As an important source of raw materials for the immune system, it participates in the synthesis of cytokines, antibodies and other immune factors; it can increase the energy level of immune cells, stimulate the proliferation and differentiation of lymphocytes, and increase the number and activity of macrophages; L-glutamine can also regulate inflammatory responses, enhance immune function and reduce the occurrence of infections.

[0041] Ascorbic acid magnesium phosphate: Combining the immune effects of vitamin C and magnesium phosphate, vitamin C can enhance the activity of immune cells, improve antioxidant capacity, regulate inflammatory response, promote the generation and activity of neutrophils, lymphocytes and macrophages, and remove free radicals in the body. As a derivative of vitamin C, magnesium phosphate can further improve the stability of vitamin C, facilitate storage and transportation, and can quickly enzymatically release vitamin C through phosphatase in the body, thereby enhancing its bioavailability in the body.

[0042] Zinc gluconate: Mainly based on the fact that zinc has the functions of immune cell production and function maintenance, regulating inflammatory response, antibacterial, participating in protein synthesis and energy metabolism, and promoting wound healing, thereby enhancing the body's immunity to pathogens.

[0043] DHA: It can directly act on immune cells, regulate immune response, maintain immune barrier, promote body health and other immune functions. DHA can promote the proliferation and differentiation of immune cells, enhance immune cell activity, promote body generation; regulate inflammatory response, enhance the synergy between immune cells; maintain intestinal mucosal barrier and skin barrier function; promote the development and function maintenance of the nervous system, remove free radicals in the body, and significantly improve the body's immunity.

[0044] EPA: can enhance the activity of immune cells, promote the phagocytosis of immune cells, and regulate the synthesis of immunoglobulins; inhibit excessive inflammatory responses and balance the functions of Th1 / Th2 cells; maintain the integrity of the intestinal mucosal barrier and enhance the defense capabilities of the skin barrier; regulate immune cell signal transduction, affect the proliferation, differentiation and apoptosis of immune cells, and thus improve the body's immunity.

[0045] Cholecalciferol: It can promote the proliferation and differentiation of immune cells such as T cells and B cells; regulate signal transduction within immune cells by binding to receptors; maintain the balance of the immune system to avoid immune diseases caused by excessive activation or inhibition of immune cells; enhance the phagocytic ability of macrophages and promote the expression of immune regulatory molecules such as cytokines and chemokines.

[0046] Based on the above principle, the present invention proposes a composition and application for enhancing immune function, including:

[0047] 1.4-1.8 parts of total ginsenosides, 12-16 parts of L-arginine hydrochloride, 18-22 parts of L-glutamine, 0.18-0.22 parts of magnesium ascorbyl phosphate, 0.02-0.06 parts of zinc gluconate, 2-6 parts of DHA, 0.6-1.0 parts of EPA, and 0.000008-0.000012 parts of cholecalciferol.

[0048] Preferably, in the above composition, the total ginsenosides are 1.6 parts, L-arginine hydrochloride is 14 parts, L-glutamine is 20 parts, ascorbic acid magnesium phosphate is 0.2 parts, zinc gluconate is 0.04 parts, DHA is 4 parts, EPA is 0.8 parts, and cholecalciferol is 0.00001 parts.

[0049] Example 1

[0050] The invention provides a preparation process of a composition for enhancing immune function.

[0051] Mainly include:

[0052] (1) Weigh the raw materials in the formula: 1.6 g of ginsenosides, 14 g of L-arginine hydrochloride, 20 g of L-glutamine, 0.2 g of magnesium ascorbyl phosphate, 0.04 g of zinc gluconate, 4 g of DHA, 0.8 g of EPA, and 0.00001 g of cholecalciferol;

[0053] (2) Mix fine powders or liquids of different raw materials together to ensure uniform composition;

[0054] (3) Dissolving the different raw materials in water and mixing them evenly to obtain a composition having an immune-enhancing function.

[0055] The above composition is used as an active ingredient and pharmaceutically acceptable excipients are added to prepare preparations, including but not limited to oral tablets, capsules, powders, chewable tablets, etc.

[0056] result:

[0057] The composition prepared by the present invention is a yellow-white suspension, which can enhance the body's immune function. The present invention proposes a scientific ratio of ginseng total saponins and other nutrients, and the formula of the composition provided by the present invention has the best immunomodulatory effect; the present invention uses an immunocompromised animal model to carry out efficacy verification of the composition formula in improving immunity; the present invention provides a composition that is more suitable for people with low immunity, which can be applied to various formula foods to achieve the effect of enhancing immunity.

[0058] Example 2

[0059] The invention provides a cellular immune function determination experiment.

[0060] Mainly include:

[0061] (1) Experimental animal culture

[0062] Healthy male Kunming mice weighing about 20g were taken, provided by Beijing Huafukang Biotechnology Co., Ltd., and free access to water and food. The animals were randomly divided into a control group (C), a cyclophosphamide model group (M), a cyclophosphamide + ginseng total saponin group (TG) and a cyclophosphamide + combination group (CP), with 10 mice in each group. The animals were adapted for 5 days before the experiment. Groups C and M were gavaged with distilled water every day, the TG group was gavaged with a ginseng total saponin solution (with water as the solvent) every day, and the CP group was gavaged with a suspension prepared with the combination (with water as the solvent) every day. On the 12th day of the experiment, mice in group C were intraperitoneally injected with normal saline, 0.2mL / mouse, and mice in groups M, TG and CP were intraperitoneally injected with 80mg / kg cyclophosphamide, 0.2mL / mouse, once a day, for 3 consecutive days.

[0063] (2) Organ / body weight ratio and cellular immune function measurement

[0064] On the 15th day of the experiment, the mice were weighed and then killed, and then soaked in 75% alcohol for 15 min. The abdominal cavity was cut open and the thymus and spleen were aseptically removed and weighed.

[0065] result:

[0066] Evaluation of the immune function of mice by spleen or thymus index:

[0067]

[0068] The spleen or thymus index was obtained according to the above formula, and Table 1 was obtained.

[0069] Table 1 Changes in spleen and thymus indexes of mice (n=10)

[0070]

[0071] a, compared with group C, P<0.05; b, compared with group M, P<0.05; c, compared with group TG, P<0.05.

[0072] C, control group; M, cyclophosphamide group; TG, total ginsenosides group; CP, combination group.

[0073] n, number of mice.

[0074] As shown in Table 1, taking ginsenosides and the composition of the present invention can significantly improve the thymus index of immunocompromised mice (P<0.05), and the composition has a better effect on improving the thymus index than ginsenosides. This indicates that the composition of the present invention can significantly enhance the organ / body weight ratio and cellular immune function.

[0075] Example 3

[0076] The invention proposes a humoral immune function determination experiment.

[0077] Mainly include:

[0078] (1) Experimental animal culture

[0079] Healthy male Kunming mice weighing about 20 g were obtained and provided by Beijing Huafukang Biotechnology Co., Ltd. They were free to drink water and eat. The animals were randomly divided into a control group (C), a cyclophosphamide model group (M), a cyclophosphamide + ginseng total saponin group (TG) and a cyclophosphamide + combination group (CP), with 5 mice in each group. The animals were adapted for 5 days before the experiment. Groups C and M were gavaged with distilled water every day, the TG group was gavaged with ginseng total saponins (with water as the solvent) every day, and the CP group was gavaged with a suspension prepared with the combination (with water as the solvent) every day. On the 12th day of the experiment, mice in group C were intraperitoneally injected with normal saline, 0.2 mL / mouse, and mice in groups M, TG and CP were intraperitoneally injected with 80 mg / kg cyclophosphamide, 0.2 mL / mouse, once a day, for 3 consecutive days; mice in group C were intraperitoneally injected with normal saline.

[0080] (2) Humoral immune function test (serum hemolysin test)

[0081] According to the half hemolysis value method (HC50 method) in the "Technical Specifications for Inspection and Evaluation of Health Foods" (2023 Edition). On the 11th day of the experiment, each mouse was intraperitoneally injected with 2% compressed SRBC 0.2mL. On the 15th day of the experiment, blood was collected and serum was separated and diluted 300 times with normal saline. Take 1mL of diluted serum and place it in a test tube, and add 10% (v / v) SRBC 0.5mL and guinea pig serum complement 1mL (diluted 1:8 with normal saline) in sequence. Set up a control tube without serum (replaced with normal saline). After keeping warm in a 37℃ constant temperature water bath for 30min, the reaction was terminated in an ice bath. Centrifuge at 2000r / min for 10min. Take 1mL of supernatant, add 3mL of Dulles reagent, and take 0.25mL of 10% (v / v) SRBC and add Dulles reagent to 4mL at the same time, mix thoroughly, let stand for 10min, use the control tube as a blank, and measure the OD value at 540nm.

[0082] result:

[0083] The humoral immune function of mice was evaluated by HC50.

[0084] The calculation of HC50 includes:

[0085] HC50 = (OD value of sample / OD value when SRBC is half hemolyzed) × dilution factor.

[0086] Table 2 Changes of serum hemolysin in mice (n=5)

[0087]

[0088] a, compared with group C, P<0.05; b, compared with group M, P<0.05; c, compared with group TG, P<0.05.

[0089] C, control group; M, cyclophosphamide group; TG, total ginsenosides group; CP, combination group.

[0090] n, number of mice.

[0091] From the results in Table 2, it can be seen that taking ginsenosides and the composition of the present invention had no significant improvement effect on serum hemolysin in immunocompromised mice (P>0.05), indicating that the composition of the present invention did not enhance immunity through humoral immune function.

[0092] Example 4

[0093] The invention provides a monocyte-macrophage function determination experiment.

[0094] Mainly include:

[0095] (1) Experimental animal culture

[0096] Healthy male Kunming mice weighing about 20 g were obtained and provided by Beijing Huafukang Biotechnology Co., Ltd. They were free to drink water and eat. The animals were randomly divided into a control group (C), a cyclophosphamide model group (M), a cyclophosphamide + ginseng total saponin group (TG) and a cyclophosphamide + combination group (CP), with 5 mice in each group. The animals were adapted for 5 days before the experiment. Groups C and M were gavaged with distilled water every day, the TG group was gavaged with ginseng total saponins (with water as the solvent) every day, and the CP group was gavaged with a suspension prepared with the combination (with water as the solvent) every day. On the 12th day of the experiment, mice in group C were intraperitoneally injected with normal saline, 0.2 mL / mouse, and mice in groups M, TG and CP were intraperitoneally injected with 80 mg / kg cyclophosphamide, 0.2 mL / mouse, once a day, for 3 consecutive days; mice in group C were intraperitoneally injected with normal saline.

[0097] (2) Monocyte-macrophage function assay (mouse peritoneal macrophage phagocytosis of chicken red blood cells)

[0098] The drop method in the "Technical Specifications for Inspection and Evaluation of Health Foods" (2023 Edition) was followed. On the 11th day of the experiment, each mouse was intraperitoneally injected with 0.2 mL of 2% compressed SRBC. On the 15th day of the experiment, the mice were killed by dislocation of the neck, and each mouse was intraperitoneally injected with 4 mL of Hank's solution containing fetal bovine serum. The abdomen was gently massaged 20 times to fully wash out the peritoneal macrophages, and then a small incision was made in the abdominal cavity, and 2 mL of peritoneal lavage fluid was drawn into the EP tube with a rubber-tipped pipette. Use a 1 mL pipette to draw 0.5 mL of peritoneal lavage fluid into a test tube containing 0.5 mL of 1% chicken red blood cell suspension and mix well. Use a syringe (with a needle) to draw 0.5 mL of the mixture and add it to the circle drawn on the slide with an immunohistochemistry pen. Place in an incubator and incubate at 37°C for 20 minutes. After the incubation, quickly rinse off the non-adherent cells with saline, fix them in methanol for 1 minute, and stain with Giemsa stain for 3 minutes. Rinse with distilled water, dry, take pictures with Olympus BX51 microscope camera system and count macrophages, calculate phagocytic rate and phagocytic index. The phagocytic rate is the percentage of macrophages that phagocytize chicken red blood cells out of every 100 macrophages; the phagocytic index is the average number of chicken red blood cells phagocytized by each macrophage.

[0099] result:

[0100] Table 3 Changes in the phagocytic percentage of mouse peritoneal macrophages (n=5)

[0101]

[0102] a, compared with group C, P<0.05; b, compared with group M, P<0.05; c, compared with group TG, P<0.05.

[0103] C, control group; M, cyclophosphamide group; TG, total ginsenosides group; CP, combination group.

[0104] n, number of mice.

[0105] As shown in Table 3, taking ginsenosides and the composition of the present invention can significantly improve the phagocytic percentage of peritoneal macrophages in immunocompromised mice (P<0.05), and the composition has a better effect on improving the phagocytic percentage of macrophages than ginsenosides. This indicates that the composition of the present invention can significantly enhance the phagocytic function of monocytes and macrophages.

[0106] Example 5

[0107] The present invention provides an NK cell activity determination experiment.

[0108] Mainly include:

[0109] (1) Experimental animal culture

[0110] Healthy male Kunming mice weighing about 20 g were obtained and provided by Beijing Huafukang Biotechnology Co., Ltd. They were free to drink water and eat. The animals were randomly divided into a control group (C), a cyclophosphamide model group (M), a cyclophosphamide + ginseng total saponin group (TG) and a cyclophosphamide + combination group (CP), with 5 mice in each group. The animals were adapted for 5 days before the experiment. Groups C and M were gavaged with distilled water every day, the TG group was gavaged with ginseng total saponins (with water as the solvent) every day, and the CP group was gavaged with a suspension prepared with the combination (with water as the solvent) every day. On the 12th day of the experiment, mice in group C were intraperitoneally injected with normal saline, 0.2 mL / mouse, and mice in groups M, TG and CP were intraperitoneally injected with 80 mg / kg cyclophosphamide, 0.2 mL / mouse, once a day, for 3 consecutive days; mice in group C were intraperitoneally injected with normal saline.

[0111] (2) NK cell activity assay

[0112] The lactate dehydrogenase activity assay method in the Technical Specifications for Inspection and Evaluation of Health Food (2023 Edition) was used. Target cells (YAC-1) were passaged 24 h before the experiment, washed three times with Hank's solution before use, and the cell concentration was adjusted to 4×10 5 The spleen was sterilely removed and ground to make a single cell suspension. It was filtered through a 200-mesh screen and centrifuged at 1000 r / min for 10 min. The supernatant was discarded, 2 mL of red blood cell lysis buffer was added and incubated at room temperature for 2 min. After centrifugation at 1000 r / min for 10 min, the cells were resuspended in 1 mL of RPMI 1640 complete culture medium containing 10% fetal bovine serum, diluted with physiological saline and counted. The final cell concentration was adjusted to 2 × 10 7 / mL. Take 100μL of target cells and effector cells, add them to a U-shaped 96-well plate; add 100μL of target cells and culture medium to the target cell natural release well, and add 100μL of target cells and 2.5% Triton to the target cell maximum release well. Set up three parallel wells for each of the above items, place in a 5% CO2, 37℃ incubator for 4h, then centrifuge the 96-well plate at 1500r / min for 10min, aspirate 100μL of supernatant from each well and place it in a flat-bottomed 96-well culture plate, add 100μL of LDH matrix solution at the same time, react at room temperature for 10min, add 30μL of 1mol / L HCl to each well, and measure the OD value at 490nm.

[0113] result:

[0114] The calculation formula is: NK cell activity (%) = (reaction well OD-natural release well OD) / (maximum release well OD-natural release well OD)×100%.

[0115] Table 4 Changes in NK cell activity in mice (n=5)

[0116]

[0117] a, compared with group C, P<0.05; b, compared with group M, P<0.05; c, compared with group TG, P<0.05.

[0118] C, control group; M, cyclophosphamide group; TG, total ginsenosides group; CP, combination group.

[0119] n, number of mice.

[0120] As shown in Table 4, taking ginsenosides and the composition of the present invention can significantly improve the NK cell activity of immunocompromised mice (P<0.05), and the composition has a better improvement effect on NK cell activity than ginsenosides, indicating that the composition of the present invention can significantly enhance NK cell activity.

[0121] Example 6

[0122] The present invention proposes a composition component selection experiment.

[0123] Mainly include:

[0124] The improvement effects of ginsenosides, ganoderma lucidum polysaccharides and astragalus polysaccharides on immunocompromised mice were compared. The dose of ginsenosides was 150 mg / kg, the dose of ganoderma lucidum polysaccharides was 120 mg / kg, and the dose of astragalus polysaccharides was 200 mg / kg. The three components were compared to improve the organ / body weight ratio, T lymphocyte transformation reaction index, and CD4+ / CD5+ in peripheral blood lymphocytes of immunocompromised mice. 3+ CD 4+Improvement effect on T cell ratio and serum immunoglobulin content.

[0125] result:

[0126] See also Figures 1 to 3 and Table 5:

[0127] Table 5 Effects of ginsenosides, ganoderma lucidum polysaccharides and astragalus polysaccharides on immune organ indexes in immunocompromised mice

[0128]

[0129] Note: *Compared with group C, P<0.05; #Compared with group M, P<0.05.

[0130] C: control group, M: immunosuppression model group, TG: ginsenosides group, GLP: Ganoderma lucidum polysaccharide group, APS: astragalus polysaccharide.

[0131] n, number of mice.

[0132] As shown in Table 1, after intraperitoneal injection of cyclophosphamide, the thymus index and spleen index of group M decreased significantly, the thymus index of mice in groups TG, GLP and APS increased compared with that in group M (P<0.05), but the spleen index of mice in groups TG, GLP and APS did not change significantly.

[0133] See also Figure 1 Cyclophosphamide significantly reduced the stimulation index of spleen T lymphocyte transformation, and TG could significantly increase the stimulation index of spleen T lymphocyte transformation in immunocompromised mice induced by cyclophosphamide (P<0.05).

[0134] See also Figure 2 Cyclophosphamide significantly reduced CD 3+ CD 4+ and CD 3+ CD 8+ T cell expression, TG and GLP can significantly increase the CD4+ T cells in peripheral blood lymphocytes of immunocompromised mice induced by cyclophosphamide. 3+ CD 4+ T cell expression, and TG has the best effect.

[0135] See also Figure 3 Cyclophosphamide significantly reduced the levels of serum IgA, IgG, and IgM. TG and GLP could significantly increase the serum IgA level in immunocompromised mice induced by cyclophosphamide, and GLP could significantly increase the IgG level (P<0.05).

[0136] Based on the above experimental data, the present invention involves adding other immune active ingredients on the basis of ginsenosides to synergistically enhance immune function. Based on experimental research, L-arginine hydrochloride, L-glutamine, magnesium ascorbyl phosphate, cholecalciferol, DHA, EPA and zinc gluconate are selected as the main raw materials. Based on the experimental results, the reasons include but are not limited to:

[0137] (1) Since L-amino acids have good biocompatibility and can be rapidly absorbed and utilized by the human body, L-arginine hydrochloride and L-glutamine were selected as one of the basic components of the formula.

[0138] (2) Compared with vitamin C, which is easily ineffective and unstable, magnesium vitamin C phosphate is a high-purity, stable derivative of vitamin C. After entering the human body, it is rapidly enzymatically hydrolyzed by phosphatase to release vitamin C, exerting the unique physiological and biochemical functions of vitamin C and avoiding the disadvantage of vitamin C being degraded before entering the human body.

[0139] (3) Cholecalciferol is the active form of vitamin D, which is more conducive to its biological function after absorption.

[0140] (4) The present invention does not use zinc sulfate or zinc carbonate in the prior art, but selects zinc gluconate which has better body absorption properties to further improve its bioavailability; based on the fact that high zinc intake can cause zinc poisoning, the dosage of zinc addition is comprehensively studied.

[0141] (5) The present invention has a significant effect on improving immune function through experiments comparing DHA / EPA=3:1 ratio. At the same time, this ratio has anti-inflammatory and antioxidant effects.

[0142] It should be noted that the term "comprises" or any other variation thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only these elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprises..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0143] The above are only embodiments of the present application and are not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included within the scope of the claims of the present application.

Claims

1. A composition for enhancing immune function, characterized in that: The components of the composition for enhancing immune function include, by weight: 1.4 to 1.8 parts of total ginsenosides, 12 to 16 parts of L-arginine hydrochloride, 18 to 22 parts of L-glutamine, 0.18 to 0.22 parts of magnesium ascorbyl phosphate, 0.02 to 0.06 parts of zinc gluconate, 2 to 6 parts of DHA, 0.6 to 1.0 parts of EPA, and 0.000008 to 0.000012 parts of cholecalciferol.

2. The composition for enhancing immune function according to claim 1, characterized in that: The total ginsenosides are 1.6 parts, the L-arginine hydrochloride is 14 parts, the L-glutamine is 20 parts, the magnesium ascorbyl phosphate is 0.2 parts, the zinc gluconate is 0.04 parts, the DHA is 4 parts, the EPA is 0.8 parts, and the cholecalciferol is 0.00001 parts.

3. Preparation of a composition for enhancing immune function, characterized in that: The composition for enhancing immune function is the composition for enhancing immune function according to claim 1 or 2, and the preparation method comprises: The ginsenosides, L-arginine hydrochloride, L-glutamine, magnesium ascorbyl phosphate, zinc gluconate, DHA, EPA and cholecalciferol are uniformly mixed.

4. A product, comprising the composition for enhancing immune function according to claim 1 or 2 and one or more additives.

5. The product according to claim 4, characterized in that The product is a pharmaceutical composition, a health product composition or a food composition.

6. The product according to claim 4, characterized in that The product is formulated to constitute tablets, capsules or granules.

7. Use of the composition for enhancing immune function according to claim 1 or 2 or the product according to any one of claims 4 to 6 in the preparation of a pharmaceutical composition for enhancing immune function of a subject.

8. The use according to claim 7, characterized in that The composition for enhancing immune function can regulate cellular immune function; The composition for enhancing immune function can enhance humoral immune function; The composition for enhancing immune function can enhance the phagocytic function of monocytes and macrophages; The composition for enhancing immune function can enhance NK cell activity.

9. Use of the composition for enhancing immune function according to claim 1 or 2 or the product according to any one of claims 4 to 6 in the preparation of a product for enhancing immune function.

10. The use according to claim 9, characterized in that: The immune-boosting products include protein powder, multivitamins and aromatherapy.

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